由农杆菌介导将白叶枯病抗性基因Xa21转入我国的5个水稻品种

利用农杆菌介导的转化系统将已克隆的Xa21基因转入我国5个水稻主栽品种, 获得了110个独立的转基因系. 转基因植株的PCR和Southern分析揭示Xa21基因已整合到受体基因组. 已整合的Xa21基因能稳定遗传, 单拷贝整合的转化体在自交T1代呈现抗感3:1的分离. 接种实验表明转基因T0植株和Xa21-PCR阳性T1植株对白叶枯病的高度抗性. 经过筛选的Xa21纯合的具有优良品质的抗性转基因系可以作为品种直接种植, 或者用于杂交稻育种.     

利用四价抗病基因提高超级杂交稻的抗性

以籼型超级杂交稻(Oryza sativa L. subsp. indica)恢复系9311的胚性愈伤组织为受体, 采用改良的基因枪轰击法将构建在同一植物表达载体上的四价抗真菌病基因(RCH10, RAC22, β-Glu和B-RIP)导入到9311的基因组中. Southern印迹杂交结果表明, 潮霉素抗性再生植株中, hpt标记基因和四价抗真菌病基因是连锁在一起并以孟德尔遗传方式进行传递的. 部分转基因R₁代和R₂代植株分别在烟溪和三亚的典型稻瘟病鉴定圃中表现出对稻瘟病菌(Magnaporthe grisea (Hebert) Barr.)的高抗性. 在高抗的转基因植株中, 多个外源基因均能正常表达. 将高抗稻瘟病的9311 R2代转基因纯系与培矮64S原种进行杂交, 杂种F₁代除对稻瘟病仍可表现出高抗性外, 还同时表现出对稻曲病和黑粉病明显提高的抗性.     

经基因工程改良的抗白叶枯病水稻粳型恢复系“C418—Xa21”及其杂交稻

通过农杆菌介导的转化系统,将业已克隆的水稻抗白叶枯病基因Xa21导入重要的粳型杂交稻恢复系“C418”。PCR和抗性分析表明单拷贝整合的Xa21在T1代的分离比为3：1。在T2代通过PCR和抗性分析选择了Xa21纯合的转基因株系“C418－Xa21”。将选择的转基因纯合系“C418－Xa21”与常用的雄性不育系“屉锦A”杂交,产生了带有转基因Xa21的杂交稻“屉优418－Xa21”（简称转基因杂交稻）。分子分析表明转基因Xa21在杂交稻“屉优418－Xa21”中能稳定遗传,抗性分析表明转基因恢复系“C418－Xa21”和转基因杂交稻“屉优418－Xa21”对白叶枯病具有高度的广谱抗性,并保持了受体对照的优良农艺性状。另外我们还转基因杂交稻“屉优418－Xa21”对白叶枯病的抗性水平高于转基因恢复系“C418－Xa21”,这可能是遗传背景的差异所致,抗白叶枯病转基因粳型恢复系数 杂交稻的育成将有益于杂交稻在我国北方稻区的推广。     

向普通小麦导入纤毛鹅观草抗黄矮病基因的研究──Ⅰ.F_1和BC_1的产生及其细胞遗传学

为了将纤毛鹅观草Z1010对黄矮病毒株系PAV和RPV的抗性基因转入普通小麦,通过幼胚拯救,获得了纤毛鹅观草Z1010×普通小麦品种莱州953的杂种F1,以及用5个普通小麦品种（系）回交的BC1衍生系。对杂种F1及BC1植株的细胞学分析表明,纤毛鹅观草Z1010不仅对Ph基因具有很强的抑制作用,而且能使杂种F1形成未减数配子,对细胞遗传学资料的进一步分析认为,通过部分同源染色体间的交换将纤毛鹅观草Z1010的抗黄矮病基因转入小麦是可能的。     

水稻白叶枯病广谱抗性基因Xa21导入两用不育系培矮64S

以克隆的Xa21基因为外源基因,成熟胚愈伤组织为转化受体,应用农杆菌介导法对水稻两用型核不育系培矮64S进行转化,获46株转基因植株。PCR和Southern分析结果表明,Xa21已整合到受体基因组。用稻白叶枯病病原菌(Xanthomonasoryzaepv.oryzae)菲律宾小种6号接种鉴定,结果表明大多数转基因植株获得了抗病性。已整合的Xa21基因能够稳定地遗传,在所检测转基因株系的T1代中,Xa21基因显示3:1的分离。     

分子标记辅助选择改良蜀恢527对白叶枯病的抗性

以含抗性基因Xa2 1和Xa4的抗白叶枯病近等基因系IRBB6 0为供体亲本 ,以不抗白叶枯病强恢复系蜀恢5 2 7为轮回受体亲本 ,连续回交 3代 ,自交 1代 ,在分离世代用分别与Xa2 1和Xa4紧密连锁的标记pTA2 48和MP12对目标基因Xa2 1和Xa4进行辅助选择 ,直至BC3F2 。在BC3F2 中选出株型、粒型、播抽期等农艺性状与蜀恢 5 2 7相似且pTA2 48和MP12的扩增带型纯合的 10个单株 ,用 10 0个RAPD和 12 0对SSR引物进行背景选择 ,决选出 5个单株 ,作为改良的蜀恢 5 2 7。抗性分析表明 ,这些改良的蜀恢 5 2 7株系对我国菌系CⅠ CⅦ和来自菲律宾的P1 和P6 均表现抗性 ,说明抗性基因已成功导入蜀恢 5 2 7中并表达。同时对pTA2 48和MP12在亲本间的多态性和选择的准确性也进行了分析 ,结果显示这两个标记在亲本间的多态性明显 ,共显性 ;选择的准确率分别在 97%和 83%以上 ,可以用其进行标记辅助选择。     

分子标记辅助选择改良水稻不育系臻达A及其杂交种的稻瘟病抗性

水稻抗稻瘟病基因Pi25是一个遗传传递能力强的广谱抗性基因。本研究以携带抗稻瘟病基因Pi25的BL27为抗源供体,与优质、配合力强、感稻瘟病的水稻保持系臻达B为受体亲本进行杂交、回交创制水稻抗病保持系新种质,再与臻达A测交和回交进行不育系转育,结合分子标记辅助选择和农艺性状筛选,获得3个抗性基因纯合、农艺性状和开花习性均与臻达A相似的改良不育系株系。利用福建省近年来致病性代表的22个稻瘟病菌株对3个改良不育系及其15个杂交种进行抗性鉴定,3个改良不育系的抗性频率为95.45%~100%,15个杂交种的抗性频率均达75%以上,而原始对照臻达A及其杂交种的抗性频率仅为54.55%和40.91%~63.64%。自然病圃诱发鉴定表明,3个改良不育系的叶瘟和穗颈瘟均为0级,表现高抗,而对照臻达A的叶瘟为5级,穗颈瘟为7级,表现感病;15个杂交种均表现良好的稻瘟病抗性。进一步分析比较15个杂交种的产量、农艺性状和稻米品质表现,结果表明臻达A-Pi25-3改良不育系的综合性状表现最优,继续回交转育,于2015年育成了稻瘟病抗性强、配合力好、群体整齐和性状稳定的不育系,命名为157A。研究表明,抗稻瘟病基因Pi25不仅在水稻不育系臻达A的遗传背景下的抗性表达完全,且在不同水稻恢复系测交种的背景下同样表现出较高水平的抗性,说明抗性基因Pi25对不育系稻瘟病改良的效果明显。创制的新不育系157A的稻瘟病抗性显著提高,还基本保留了原来不育系高配合力等优良特性,为选育高产、优质、抗病杂交稻新品种提供了不育系新种质。     

三基因聚合改良恢复系福恢673的稻瘟病抗性

为了改良水稻优良恢复系福恢673的稻瘟病抗性,以该恢复系为轮回亲本,以携有3个稻瘟病抗性基因(Pi-1、Pi-9和Pi-k~h)的优质恢复系金恢1059为供体亲本,通过回交育种结合分子标记辅助选择,选育出10个导入了这3个抗稻瘟病基因的福恢673近等基因系,其遗传背景恢复率为92.96%–98.59%。抗性鉴定结果表明,这些近等基因系及其与不育系宜香A配制的杂种一代均表现抗稻瘟病,抗性明显强于对照福恢673和宜优673,且半数以上杂种一代基本保留了原组合的主要优点。用近等基因系Line 9配组的杂交稻新组合两优7283和金泰优683在区试中均表现出产量高、稻瘟病抗性强、生育期适中等特点,表明该近等基因系具较好的应用前景。     

多种优质高分子量谷蛋白亚基的聚合育种研究

用携带HMW-GS14 15的小偃6号作为轮回亲本,携带HMW-GS5 10的法国优质面包小麦品系707作为供体亲本,在回交后代的BC1、BC2、BC3、BC3F1、BC3F2代其它农艺性状选择的基础上,利用1对特异引物逐代检测出携带优质,Dx5基因的单株进行回交和自交。BC1代中随机检测的58个单株的Dx5基因分布符合1对等位基因的遗传分离比例1：1;BC1代小麦相同3个单株3个不同生长季节Dx5基因检测的结果完全一致,检测结果非常稳定;已将优质Dx5基因导入BC3F2后代的部分单株内;携带Dx5基因的株系XN89-7-3微量SDS沉淀值为18．8mL,比小偃6号提高了23．68％;蛋白质电泳筛选出了6个聚合多种优质亚基且编码基因纯合的单株,微量SDS沉淀值为19．9mL,比小偃6号提高了30．92％;选择农艺性状与轮回亲本相似并具有Dx5基因特异扩增产物的单株进行回交或自交,可加快回交转育的进度。实践证明,回交转育与分子标记辅助选育相结合的育种方法是快速定向聚合多种优质HMW-GS基因的有效方法之一。     

无选择标记和载体骨干序列的Xa21转基因水稻的获得

利用双右边界T-DNA载体通过根癌农杆菌介导法将水稻白叶枯病广谱抗性基因Xa21导入杂交稻重要恢复系C418中。T0代共获得27个独立转基因株系,通过田间抗性鉴定与PCR分析,有17个株系的Xa21基因分子鉴定为阳性,且对白叶枯病原菌P6生理小种具有抗性。通过对17个株系的后代植株进行田间抗性鉴定,分子标记辅助选择及Southern杂交分析,结果显示4个株系的T1代植株中能分离出无潮霉素标记基因的Xa21转基因植株。无选择标记Xa21转基因株系的获得率为15%。PCR检测还表明,这些无选择标记的Xa21转基因植株不带有载体骨架序列。通过对转基因后代进一步的抗性鉴定与PCR辅助选择,获得了无选择标记和载体骨架序列的转基因Xa21纯合的抗白叶枯病水稻。     

我国麦蚜抗药性及研究现状

麦蚜是危害我国小麦Triticum aestivum L.生产的主要害虫,具有分布广、数量大、繁殖力强以及远距离迁飞等特点,不仅直接吸食小麦汁液,还传播多种植物病毒,每年造成小麦减产10%~30%。目前对麦蚜的防控主要以化学防治为主,但由于化学杀虫剂长期或不合理的使用,多地麦蚜对常用杀虫剂产生了不同程度的抗性。本文从麦蚜抗药性测定方法、抗性水平及交互抗性、代谢和靶标抗性机制、以及麦蚜抗药性综合治理等方面进行了综述,以期为麦蚜的防治及杀虫剂的持续合理使用提供理论参考和依据。     

Stability analyses for estimating relative durability of quantitative resistance

Summary Experiments in which a series of host cultivars are inoculated in all combinations with a series of pathogen isolates have been used to detect specificity in the host resistance. A theoretical model of polygenic resistance involving both general and specific interactions with pathogen virulence was developed to test the abilities of statistical analyses to discriminate between host genotypes with different levels of general and specific resistance. Estimates of levels of specific resistance could be obtained in regressions of disease severity scores for each host cultivar X pathogen isolate combination vs. the virulence index of each isolate. If the virulence index was based on the mean disease severity induced by the isolate over all host cultivars, the slopes of the regression lines were correlated with the levels of specific resistance in host cultivars. If the virulence index was based on the disease severity induced by the isolate on a host cultivar with a minimum of specific resistance, the mean squares for deviations from the regression were correlated with the levels of specific resistance in host cultivars. A method was developed to consistently choose host cultivars with minimum specific resistance. The two regression analyses gave estimates of specificity in randomly generated, model genotypes of approximately equal accuracy, although the second method appeared to be more accurate when the numbers of loci controlling resistance and virulence were small. The best estimates of numbers of genes for specific resistance were obtained by calculating a rating based on mean disease severity, the mean square for deviation from the regression on the virulence index based on disease severity on the cultivar with minimum specific resistance and the slope of the regression on the virulence index based on the mean disease severity. The best estimates of proportions of resistance genes that were specific were obtained by calculating a rating based on the above deviation mean square and slope alone.Cooperative investigation of the U.S. Department of Agriculture, Agricultural Research Service and the North Carolina Agricultural Research Service. Journal Series Paper No. 8326 of the North Carolina Agricultural Research Service     

西花蓟马的抗药性及其治理策略

西花蓟马Frankliniella occidentalis（Pergande）是世界性的园艺作物上的重要害虫,几乎对每种类型的杀虫剂均产生了抗药性,包括有机磷、氨基甲酸酯、拟除虫菊酯类杀虫剂和多杀菌素等。本文对国外西花蓟马的抗药性发展现状和抗性机制进行了总结,并提出了抗性治理策略,即科学合理使用杀虫剂,结合栽培防治、物理防治、生物防治和寄主植物抗性等方式降低杀虫剂对西花蓟马的选择压,从而达到抗性治理的目的。     

PC12 Variants Deficient in Catecholamine Transport

We have isolated PC12 cell variants deficient in transporter-mediated uptake of 3,4-dihydroxyphenylethylamine (dopamine). The variants either were obtained nonselectively, or they were selected by resistance to guanethidine or N-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP). Dopamine uptake into guanethidine-resistant cells occurred with a decreased Vmax; the Km for dopamine and inhibition by guanethidine were normal. MPTP-resistant cells lacked the capacity to take up dopamine. Most of the variants resembled wild-type PC12 in their response to nerve growth factor and the storage and secretion of dopamine. MPTP-resistant cells exhibited several deficiencies in addition to dopamine transport, i.e., no measurable storage of dopamine or acetylcholine and no observable response to nerve growth factor. Wild-type and variant cells were compared with respect to the labeling of cell proteins with [3H]xylamine, which binds covalently to certain proteins apparently only after entering PC12 via the catecholamine transporter. When intact variant cells were used, there was markedly reduced labeling of the proteins by [3H]xylamine. Almost all of these proteins were readily labeled when cell homogenates were exposed to [3H]xylamine. However, MPTP-resistant cells were missing three of these proteins. Northern blot analysis with cDNA clones revealed that the MPTP-resistant cells had markedly reduced levels of several specific mRNA species.     

甜菜夜蛾的抗性及抗性机理研究进展

甜菜夜蛾Spodoptera exigua(Hübner)是世界性农业害虫,近年来在我国由次要害虫上升为主要害虫。甜菜夜蛾对很多种化学杀虫剂和生物杀虫剂产生了抗性。本文分别阐述了甜菜夜蛾对常用杀虫剂如有机磷类、拟除虫菊酯类、氨基甲酸酯类、生长调节类杀虫剂、Bt杀虫剂等的抗性发展现状,并且对抗药性机理进行了总结,提出了抗性治理措施。     

Selection of a dexamethasone-resistant H-4-IIE-C3 rat hepatoma tissue-culture line

Summary Exposure of H-4-IIE-C3 rat hepatoma cell cultures to the synthetic glucocorticoid, dexamethasone, results in an inhibition of cellular proliferation which is not the result of steroid-induced cytolysis. A significant decrease in both the rat of DNA synthesis and DNA content precedes, a detectable effect on cell number. Continuous culture of H-4-IIE-C3 cells in medium containing 10⁻⁵ m dexamethasone results in the selection of a steroid-resistant cell population that has the growth characteristics of unselected sensitive cultures and shows normal steroid induction of tyrosine transaminase. Selection is a slow process requiring 24 to 36 months to obtain, a phenotypically stable resitant cell line, and can be subdivided into three phases—a sensitive phase, adaptation and resistance. A comparison of the karyotypes of unselected and resistant cultures shows that the selection process enriches for a dexamethasone-resistant subpopulation. This work was supported by Special Grant No. 716 from the California Division of the American Cancer Society, and a grant-in-aid from the San Diego State University Foundation.     

Herbicide resistance—what have we learned from other disciplines?

Herbicide resistance is a growing threat to agriculture and has parallels to resistances to fungicides and insecticides. However, there are many reasons to treat the resistance to herbicides differently. To highlight these similarities and differences, three pests, a weed, an insect, and a disease that have shown the ability to rapidly develop resistance to a variety of products and product classes were used as illustrations. The situation in herbicide resistance is approaching a point already experienced by the other pest control disciplines, and thus, it is worthwhile to revisit their experiences.     

种子内生菌增强宿主植物重金属抗性的功能机制研究进展

种子是植物的繁殖器官,其内定殖有一定数量的内生菌,种子内生菌通过垂直传播成为新生植物组织内最早定殖的微生物,对连续几代植物内生菌群落的形成起着决定性作用,并在植物抗逆方面发挥着重要作用。本文对种子内生菌与宿主植物重金属抗性之间的关系及其功能机制进行综述,并对下一步研究方向予以展望。